Associação de Fatores de Risco Cardiovascular e Polimorfismo de APOE com Mortalidade em Idosos Longevos: Uma Coorte de 21 Anos / Association of Cardiovascular Risk Factors and APOE Polymorphism with Mortality in the Oldest Old: A 21-Year Cohort Study

Resumo Fundamento: O conhecimento dos fatores ambientais e genéticos para um envelhecimento bem-sucedido em idosos longevos é controverso. Acrescenta-se a esta evidência, o fato de serem poucos os estudos delineados com essa população. Objetivo: Investigar a relação entre os genótipos mais frequentes da apolipoproteína E (APOE) e a mortalidade em idosos longevos que vivem em comunidade e sua sobrevida de acordo com os fatores de risco cardiovascular. Métodos: Uma amostra de 74 idosos com 80 anos ou mais da coorte do Projeto Veranópolis foi selecionada para genotipagem da APOE. Na linha de base, foram coletadas variáveis antropométricas, dosagens sanguíneas de glicose e lipídeos, pressão arterial e variáveis de estilo de vida (tabagismo, consumo de álcool e atividade física). A escala Bayer de Atividades da Vida Diária foi aplicada aos cuidadores dos idosos. O tempo de seguimento total do estudo foi 21 anos. Um p<0,05 bicaudal foi considerado estatisticamente significativo. Resultados: Não encontramos associação entre os genótipos da APOE e mortalidade. Entretanto, o risco de morte em idosos fumantes foi 2,30 vezes (hazard ratio [HR]; intervalo de confiança de 95% [IC 95%] 1,01 a 5,24); em diabéticos, 3,95 vezes (HR; IC 95% 1,27 a 12,30) do risco dos não diabéticos. Indivíduos que praticavam atividade física vigorosa tiveram uma redução no risco de óbito em 51% (HR = 0,49; IC 95% 0,27 a 0,88). Para o aumento de 1 mmHg na pressão arterial sistólica houve uma redução de 2% (HR = 0,98; IC 95% 0,97 a 0,99) no risco de morte. Conclusão: Nesta amostra de longevos, não houve associação entre os genótipos da APOE e mortalidade. Entretanto, os fatores de risco cardiovasculares clássicos podem ser importantes para a mortalidade geral em pessoas muito idosas.

Abstract Background: Knowledge of environmental and genetic factors for healthy aging in elderly people is controversial. In addition to this evidence, few studies have been designed for this population. Objectives: To investigate the relationship between the most frequent apolipoprotein E (APOE) genotypes and mortality in very elderly individuals living in a community and to evaluate survival according to cardiovascular risk factors. Methods: A sample of 74 elderly individuals aged ≥ 80 years, from the Veranópolis Project cohort, was selected for APOE genotyping. At baseline, anthropometric variables, glucose and lipid levels, blood pressure, and lifestyle variables (smoking, alcohol consumption, and physical activity) were collected. The Bayer Activities of Daily Living Scale was applied to their caregivers. Total study follow-up was 21 years. Two-sided p < 0.05 was considered statistically significant. Results: There was no association between APOE genotypes and mortality. However, the risk of death in elderly smokers was 2.30 times higher (hazard ratio [HR], 95% CI 1.01 to 5.24); in individuals with diabetes, it was 3.95 times higher (HR, 95% CI 1.27 to 12.30) than in individuals without diabetes. Subjects who practiced vigorous physical activity had a 51% reduction in risk of death (HR = 0.49, 95% CI 0.27 to 0.88). For an increase of 1 mmHg in systolic blood pressure, there was a 2% reduction (HR = 0.98, 95% CI 0.97 to 0.99) in risk of death. Conclusion: In this sample population, APOE genotypes were not associated with mortality. However, classic cardiovascular risk factors may be important for overall mortality in the very elderly.

Association of Cardiovascular Risk Factors and APOE Polymorphism with Mortality in the Oldest Old: A 21-Year Cohort Study. / Associação de Fatores de Risco Cardiovascular e Polimorfismo de APOE com Mortalidade em Idosos Longevos: Uma Coorte de 21 Anos.

BACKGROUND: Knowledge of environmental and genetic factors for healthy aging in elderly people is controversial. In addition to this evidence, few studies have been designed for this population. OBJECTIVES: To investigate the relationship between the most frequent apolipoprotein E (APOE) genotypes and mortality in very elderly individuals living in a community and to evaluate survival according to cardiovascular risk factors. METHODS: A sample of 74 elderly individuals aged ≥ 80 years, from the Veranópolis Project cohort, was selected for APOE genotyping. At baseline, anthropometric variables, glucose and lipid levels, blood pressure, and lifestyle variables (smoking, alcohol consumption, and physical activity) were collected. The Bayer Activities of Daily Living Scale was applied to their caregivers. Total study follow-up was 21 years. Two-sided p < 0.05 was considered statistically significant. RESULTS: There was no association between APOE genotypes and mortality. However, the risk of death in elderly smokers was 2.30 times higher (hazard ratio [HR], 95% CI 1.01 to 5.24); in individuals with diabetes, it was 3.95 times higher (HR, 95% CI 1.27 to 12.30) than in individuals without diabetes. Subjects who practiced vigorous physical activity had a 51% reduction in risk of death (HR = 0.49, 95% CI 0.27 to 0.88). For an increase of 1 mmHg in systolic blood pressure, there was a 2% reduction (HR = 0.98, 95% CI 0.97 to 0.99) in risk of death. CONCLUSION: In this sample population, APOE genotypes were not associated with mortality. However, classic cardiovascular risk factors may be important for overall mortality in the very elderly.

FUNDAMENTO: O conhecimento dos fatores ambientais e genéticos para um envelhecimento bem-sucedido em idosos longevos é controverso. Acrescenta-se a esta evidência, o fato de serem poucos os estudos delineados com essa população. OBJETIVO: Investigar a relação entre os genótipos mais frequentes da apolipoproteína E (APOE) e a mortalidade em idosos longevos que vivem em comunidade e sua sobrevida de acordo com os fatores de risco cardiovascular. MÉTODOS: Uma amostra de 74 idosos com 80 anos ou mais da coorte do Projeto Veranópolis foi selecionada para genotipagem da APOE. Na linha de base, foram coletadas variáveis antropométricas, dosagens sanguíneas de glicose e lipídeos, pressão arterial e variáveis de estilo de vida (tabagismo, consumo de álcool e atividade física). A escala Bayer de Atividades da Vida Diária foi aplicada aos cuidadores dos idosos. O tempo de seguimento total do estudo foi 21 anos. Um p<0,05 bicaudal foi considerado estatisticamente significativo. RESULTADOS: Não encontramos associação entre os genótipos da APOE e mortalidade. Entretanto, o risco de morte em idosos fumantes foi 2,30 vezes (hazard ratio [HR]; intervalo de confiança de 95% [IC 95%] 1,01 a 5,24); em diabéticos, 3,95 vezes (HR; IC 95% 1,27 a 12,30) do risco dos não diabéticos. Indivíduos que praticavam atividade física vigorosa tiveram uma redução no risco de óbito em 51% (HR = 0,49; IC 95% 0,27 a 0,88). Para o aumento de 1 mmHg na pressão arterial sistólica houve uma redução de 2% (HR = 0,98; IC 95% 0,97 a 0,99) no risco de morte. CONCLUSÃO: Nesta amostra de longevos, não houve associação entre os genótipos da APOE e mortalidade. Entretanto, os fatores de risco cardiovasculares clássicos podem ser importantes para a mortalidade geral em pessoas muito idosas.

The Brazilian Cardioprotective Nutritional Program to reduce events and risk factors in secondary prevention for cardiovascular disease: study protocol (The BALANCE Program Trial).

This article reports the rationale for the Brazilian Cardioprotective Nutritional Program (BALANCE Program) Trial. This pragmatic, multicenter, nationwide, randomized, concealed, controlled trial was designed to investigate the effects of the BALANCE Program in reducing cardiovascular events. The BALANCE Program consists of a prescribed diet guided by nutritional content recommendations from Brazilian national guidelines using a unique nutritional education strategy, which includes suggestions of affordable foods. In addition, the Program focuses on intensive follow-up through one-on-one visits, group sessions, and phone calls. In this trial, participants 45 years or older with any evidence of established cardiovascular disease will be randomized to the BALANCE or control groups. Those in the BALANCE group will receive the afore mentioned program interventions, while controls will be given generic advice on how to follow a low-fat, low-energy, low-sodium, and low-cholesterol diet, with a view to achieving Brazilian nutritional guideline recommendations. The primary outcome is a composite of death (any cause), cardiac arrest, acute myocardial infarction, stroke, myocardial revascularization, amputation for peripheral arterial disease, or hospitalization for unstable angina. A total of 2468 patients will be enrolled in 34 sites and followed up for up to 48 months. If the BALANCE Program is found to decrease cardiovascular events and reduce risk factors, this may represent an advance in the care of patients with cardiovascular disease.

Alpha-keto-beta-methylvaleric acid increases the in vitro phosphorylation of intermediate filaments in cerebral cortex of young rats through the gabaergic system.

In this study we investigated the effects of alpha-ketoisovaleric (KIV) and alpha-keto-beta-methylvaleric acids (KMV), metabolites accumulating in the inherited neurometabolic disorder maple syrup urine disease (MSUD), on the in vitro incorporation of 32P into intermediate filament (IF) proteins from cerebral cortex of young rats during development (9-21 days of age) We observed that KMV significantly increased the in vitro incorporation of 32P into the IF proteins studied in cortical slices of 12-day-old rats through the PKA and PKCaMII, with no alteration at the other ages. In contrast, KIV was ineffective in altering the phosphorylating system associated with IF proteins at all ages examined. A similar effect on IF phosphorylation was achieved by incubating cortical slices with gamma-aminobutiric acid (GABA). Furthermore, by using specific GABA antagonists, we verified that KMV induced a stimulatory effect on IF phosphorylation of tissue slices from 12-day-old rats mediated by GABA(A) and GABA(B) receptors. In conclusion, our results indicate the involvement of the GABAergic system in the alterations of IF phosphorylation caused by KMV, one of the branched-chain keto acids accumulating in MSUD.

Effect of propionic and methylmalonic acids on the in vitro phosphorylation of intermediate filaments from cerebral cortex of rats during development.

In this study we investigated the in vivo and in vitro effects of methylmalonic (MMA) and propionic acids (PA), at concentrations usually found in methylmalonic acidemia and propionic acidemia respectively, on the phosphorylation of intermediate filament proteins in cerebral cortex of rats during development. Rats of 9, 12, and 17 days were acutely injected with the acids and sacrificed 90 min after injection. The cerebral cortex was dissected, and slices were incubated with 32P-orthophosphate. The cytoskeletal fraction was extracted and the radioactivity incorporated into intermediate filament subunits was measured. In addition, cortical slices from nontreated rats of 9, 12, 15, 17, 21, and 60 days of life were incubated with the acids in the presence of 32P-orthophosphate, the cytoskeletal fraction was extracted and the radioactivity was measured. Results demonstrated that MMA and PA significantly decreased the radioactivity incorporated into intermediate filament proteins at day 12, both in vivo and in tissue slices. In contrast, PA increased the in vitro phosphorylation of the cytoskeletal proteins in slices of 21-day-old animals. It acts through PP2A and PP2B in 12-day-old rats and through PKA and PKCaMII in 21-day-old animals. We propose that alteration of cytoskeletal protein phosphorylation caused by methylmalonic and propionic acids may be related to the neurological dysfunction characteristic of propionic and methylmalonic acidemia.

In vitro phosphorylation of cytoskeletal proteins from cerebral cortex of rats.

Procedures for the preparation of high- and low-salt Triton insoluble cytoskeletal fractions from rat brain suitable for studying in vitro phosphorylation by endogenous kinases and phosphatases are described. The high-salt Triton insoluble cytoskeletal fraction is enriched in neurofilament subunits (NF-H, NF-M and NF-L), vimentin and glial fibrillary acidic protein (GFAP), while the low-salt Triton insoluble cytoskeletal fraction contains detergent insoluble cytoskeletal elements such as intermediate filament subunits and tubulins. One of our approaches is to incubate cerebral cortex slices with [32P]orthophosphate before the cytoskeletal fraction extraction, which allows the in vitro phosphorylation of cytoskeletal constituents in an intact intracellular environment. On the other hand, we also incubate low- or high-salt cytoskeletal fractions previously prepared with [gamma(32)P]ATP. By doing so, we are able to study the direct effects of substances on the kinase and phosphatase activities associated with the cytoskeletal fraction. Moreover by using specific activators or inhibitors of protein kinases and phosphatases we can obtain more detailed information on the alterations provoked by these substances. These approaches are useful for the investigation of the neurotoxic effects of various drugs and metabolites affecting the cytoskeletal-associated phosphorylation system in the brain.

alpha-Ketoisocaproic acid regulates phosphorylation of intermediate filaments in postnatal rat cortical slices through ionotropic glutamatergic receptors.

In this study we investigated the effects of alpha-ketoisocaproic acid (KIC), the main keto acid accumulating in the inherited neurometabolic disorder maple syrup urine disease (MSUD), on the in vitro incorporation of 32P into intermediate filament (IF) proteins from cerebral cortex of rats during development. KIC decreased the in vitro incorporation of 32P into the IF proteins studied up to day 12, had no effect on day 15, and increased this phosphorylation in cortical slices of 17- and 21-day-old rats. A similar effect on IF phosphorylation was achieved along development by incubating cortical slices with glutamate. Furthermore, the altered phosphorylation caused by the presence of KIC in the incubation medium was mediated by the ionotropic receptors NMDA, AMPA and kainate up to day 12 and by NMDA and AMPA in tissue slices from 17- and 21-day-old rats. The results suggest that alterations of IF phosphorylation may be associated with the neuropathology of MSUD.

Cytoskeleton of human mononuclear cells as a possible peripheral marker for phenylalanine neurotoxicity in PKU.

In this work we tested human mononuclear cells as a peripheral marker to study neurotoxicity of phenylalanine (Phe). Slices of cerebral cortex of rats or human mononuclear cells were incubated with different concentrations of Phe and/or Ala in the presence of 32P-orthophosphate, the cytoskeletal fraction was extracted, and the radioactivity incorporated into intermediate filament proteins was measured. Our results show that 2 mM Phe as well as 1 mM Ala are effective in increasing the 32P in vitro incorporation into IFs in both tissues. When cerebral cortex slices or mononuclear cells were incubated with different concentrations of Phe and/or Ala, the effects on the 32P in vitro incorporation into IF proteins was compatible with an antagonistic mechanism of action of the two amino acids on the enzymes of the phosphorylating system. In addition, these blood cells may be a possible peripheral marker to study neurotoxicity of Phe in patients with PKU.